Valorization of three varieties of grape

In the present investigation, seed methanolic extracts of three Vitis vinifera (Muscat d’Italie, Syrah and Carignan) were assayed for their antioxidant activities. Results showed that there are strong variations in the contents of total phenols (440.97–121.94 mg GAE g⁻¹ DW), flavonoids (48.07–16.81 mg EC g⁻¹ DW) and tanins (37.15–14.9 mg EC g⁻¹ DW) from the studied seeds. The phenolic composition of these extracts was determined by RP–HPLC after acid hydrolysis. The main phenolic compound was quercetin with 27.2% in Muscat d’Italie, 48.8% in Syrah and 28.4% in Carignan. Besides, all seed extracts showed remarkable DPPH radical scavenging activity with IC₅₀ values ranged from 1.8 to 30 μg ml⁻¹. EC₅₀ values of reducing power activity ranged from 100 to 120 μg ml⁻¹. The high phenolic content and the considerable antioxidant activities of vitis seed extracts could potentially be considered as an expensive source of natural antioxidants.     

Growing conditions and postharvest management can affect the essential oil of Origanum vulgare L. ssp. hirtum (Link) Ietswaart

Origanum vulgare subsp. hirtum (Link) Ietswaart (Greek oregano) has been cultivated since ancient times thanks to its herbal and therapeutic properties. Currently, it is added in various commercial preparations thanks to its essential oil composition. This group of secondary compounds is affected in quantity and quality by biotic and abiotic factors during the cultivating phase and by the postharvest management. Thus, the aims of the work were to study: (1) how the growing conditions (soil full-light treatment; pot full-light treatment; pot 50%-shade treatment) can affect the essential oil content and the essential oil profile of Greek oregano branches at full-blossoming stage; and (2) how the postharvest management (distillation of the essential oil from fresh herbs, dehumidified herbs or oven-dried herbs) can affect the essential oil content and the essential oil profile of this species. The growing conditions significantly affected the biometrical parameters. The pot full-light treatment contained the highest dry matter percentage (36.5%) and the highest essential oil content, which was directly correlated to the former parameter (r = 0.890**). The postharvest management did not affect the essential oil content. The growing condition significantly affected the characterizing γ-terpinene, cis-sabinene hydrate, 4-terpineol, α-terpinene, linalyl acetate, and β-bisabolene, while it did not affect carvacrol (mean value 15.8%), p-cymene (mean value 6.0%), or sabinene (mean value 3.9%). The postharvest management significantly affected carvacrol, cis-sabinene hydrate, 4-terpineol, p-cymene, α-terpinene, linalyl acetate, and β-bisabolene, while it did not affect γ-terpinene (mean value 14.7%). The response of some compounds to growing conditions led us to hypothesise an essential oil profile of the plants grown in pots at 50%-shade to be more of a vegetative stage type oil than the essential oil profile of the plants grown in soil, although all were at the blossoming stage. The analyses on the individual compounds indicated that, even though the essential oil content in oregano did not change with a change in postharvest management, biochemical changes occurred on the profile. These could be due to enzyme activity, dilution effect or temperature effect during the drying processes. The oven-drying process and the dehumidifying process changed the essential oil profile in a similar way compared to the essential oil profile distilled from fresh herbs. Thus, when it is not possible to extract the essential oil directly from fresh herbs, processors can dry the oregano according to the equipments they have available.     

Influence of chemical and biological treatments on volatile oil composition of Artemisia annua Linn.

The relative abundance of isoprenoids and other volatile components in the aerial parts of Artemisia annua Linn. (Asteraceae) after different nutritional treatments was analyzed by GC and GC/MS. Hydro-distillation of untreated (control) plants yielded 0.28 ± 0.04% essential oil on fresh weight basis. Monoterpenes were predominant in the essential oil of which cis-limonene-1,2-epoxide (22.1%), artemisia ketone (11.5%), iso-pinocamphone (11.4%), thujyl alcohol (9.9%) and camphor (8.4%) were the major components. The essential oil distilled from A. annua treated with Azospirillum (2 ml/plant) amounted to 0.30 ± 0.03% of fresh weight and consisted mainly of neral (31.1%), β-caryophyllene (25.1%), artemisia ketone (10.0%), thujyl alcohol (9.4%), trans-bergamotene (8.5%) and spathulenol (4.8%). The essential oil from plants treated with basal N, P, K and S application (in the form of urea P₂O₅, K₂O and gypsum at a rate of 120, 50, 100 and 50 kg/ha, respectively) amounted to 0.32 ± 0.03% of fresh weight and consisted mainly of thujyl alcohol (33.3%), β-caryophyllene (15.5%), cis-undec-5-ene (14.4%), artemisia ketone (6.0%), trans-nerolidol (5.8%) and undec-4-ene (4.6%). The hydrodistillation of A. annua treated with Glomus (Mycorrhizal soil 50 g/plant) yielded 0.50 ± 0.02% essential oil on fresh weight basis. The sesquiterpenes were predominant in this essential oil and consisted of β-caryophyllene (51.2%), trans-bergamotene (15.4%), α-gurjunene (14.3%), germacrene D (5.1%) and ledol (4.5%) as major components. The only monoterpenes in the essential oil were neral, nerol and pulegone (each less than 1%). These findings suggest that A. annua treated with Glomus lacks glandular trichomes—the primary sites of synthesis of monoterpenes.     

Prior weakening of Macrophomina phaseolina and Fusarium propagules for enhancing efficiency of Brassica amendments

In a two year field study, the effect of varying intensities of sub-lethal heating on the efficiency of Brassica amendments in controlling viable populations of Macrophomina phaseolina and Fusarium oxysporum f sp. cumini was determined in an arid region of India. After 30 d of dry summer exposure of pathogen infested soil, incorporation of mustard residues and oil cake (0.18% and 0.04% w/w) and then applying one irrigation caused significant reduction by 75.3–81.3% in viable counts of M. phaseolina that causes dry root rot of legumes and by 93.9% in counts of F.o. f. sp. cumini causing wilt of cumin (Cuminum cyminum L.) at 0–15 and 16–30 cm depths. Increasing duration of summer exposure to 60 d improved the reductions in viable propagules of M. phaseolina by 83.6–90.4% and in F.o. f. sp. cumini by 78.2–94.8% at same soil depths. At certain heat levels, reduction in viable population of Fusarium due to amendments and irrigation was greater than that recorded in Macrophomina. Significantly low levels of reduction in pathogenic propagules of Macrophomina (63.9–71.4%) and Fusarium (48.0–57.2%) under shade compared to unshaded conditions indicated that mild heating did not cause discernible weakening effect. In second season also, 89.2–91.5% and 78.5–95.8% reduction in counts of Macrophomina and Fusarium, respectively was achieved by the application of amendments after 60 d of summer exposure at 0–30 cm soil depth. These results suggested a new approach to improve the control of soil-borne plant pathogens in hot arid regions by combining prolonged sub-lethal heating, effective naturally available on-farm wastes as soil amendments and one summer irrigation.     

Nutrient composition of the leaves and flowers ofColocasia esculenta and the fruits ofSolanum melongena

The nutrient composition, ofColocasia esculenta flowers (CF) and leaves (CL), and the green fruits ofSolanum melongena (SM) were carried out as a means to determine their nutritional potential. Results showed that these food materials had high moisture and fiber levels which ranged between 888 and 906 g·kg^–1; and 204 and 303 g·kg^–1 dry weight (dw) for moisture and fiber respectively. The calorific values were between 3889 and 4001 kcals·kg^–1 dw, while the total lipids ranged from 53 in CF to 71 g·kg^–1 dw in SM. The leaves ofColocasia esculenta had the highest crude protein value of 307 g·kg^–1 dw. The flowers had 149 g·kg^–1 dw while SM had 138 g·kg^–1 dw. The amino acid profile in the flowers and leaves ofColocasia esculenta in contrast to SM were balanced comparable to the reference FAO pattern. Ash values were high (ranging from 76 in SM to 98 g·kg^–1 in CL) with potassium being the principal element. Iron and Zinc levels were also high especially in CF (with 303 and 82 mg·kg^–1 dw respectively). These foods also contained moderate quantities of calcium, phosphorus and magnesium but were poor in manganese and copper.     

Agrobotanical traits and chemical composition of Cassia obtusifolia L.: A lesser-known legume of the Western Ghats region of South India

Four accessions of the lesser-known legume, Cassia obtusifolia L. (Sickle pod), collected from four different agroclimatic regions of Western Ghats, were evaluated for agrobotanical traits and chemical composition. Amongthe four accessions, the Keriparai accession had the highest values for plant height (cm), number of flowers per cluster, number of pods per cluster, pod length (cm), seeds per pod, seed weight (g) per pod and seed recovery percentage. Crude protein ranged from 18.56–22.93%, crude lipid was between 5.35–7.40%, crude fiber ranged from 6.83–9.45%, ash content ranged from 5.14–5.83% and carbohydrate varied from 57.00–60.69%. Globulins constituted the bulk of the seed protein as in most legumes. Mineral profiles, viz., sodium, potassium, calcium, magnesium, phosphorus, iron, copper, zinc and manganese ranged from 42.92–84.83, 758.05–1555.79, 559.92–791.72, 456.36–709.47, 629.13–947.79, 8.42–12.35, 0.93–2.06, 10.60–30.04 and 2.12–4.12 Mg/100 g seeds flour, respectively. Seed proteins of all accessions exhibited relatively high levels of non-essential and essential amino acids, with the exception of threonine. The in vitro protein digestibility of the legume ranged from 74.66 to 81.44%. Antinutritional substances such as total free phenolics ranged from 0.34–0.66%; tannins were between 0.47–0.60%, L-DOPA content ranged from 0.98–1.34%; trypsin inhibitor activity varied from 11.4–13.5 TIU/mg protein and chymotrypsin inhibitor activity ranged from 10.8–12.3 CIU/mg protein. Phytohemagglutinating activity also was assayed. Inconclusion, the accessions of C. obtusifolia, collected from Western Ghats, South India, could serve as a low-cost source of some important nutrients for humans. The antinutritional factors might have little nutritional significance, if the seeds are processed properly.     

Chemical composition of the underutilized legume Cassia hirsuta L.

Seven accessions of the underutilized legume, Cassia hirsuta L., seeds collected from seven different agroclimatic regions of Tamil Nadu, India, were analyzed for proximate composition, total proteins, protein fractions, mineral profiles and selected antinutritional factors. Crude protein ranged from 15.52 to 20.74%, crude lipid 3.77–7.04%, crude fiber4.68–6.92%, ash 3.98–6.42% and carbohydrates 62.45–70.16%. Energyvalues of the seeds were 1549–1634 kJ/100 g (DM), which are comparable to those of other legumes. Data on seed protein fractions revealed thatglobulins constituted the bulk of the seed protein as in most legumes. Mineralcontents of the seeds showed greater variation. Potassium was the mostabundant mineral (1029–1786 mg/100 g), whereas manganese was low (2.1–2.2 mg/100 g). Antinutritional factors such as total free phenolics,tannins, L-DOPA and lectins were analyzed. The results of the studydemonstrated that the accessions of C. hirsuta seeds collectedfrom Tamil Nadu, India, could be good sources of some important nutrientsfor humans.     

Essential oil composition of Hypericum perfoliatum L. and Hypericum tomentosum L. growing wild in Tunisia

The essential oils obtained by hydrodistillation from the aerial parts of Tunisian native Hypericum perfoliatum L. (sect. Drosocarpium Spach.) and Hypericum tomentosum (sect. Adenosepalum Spach.) were analyzed by GC and GC–MS. Thirty-two compounds were identified in the essential oils of H. perfoliatum with α-pinene (13.1%), allo-aromadendrene (11.4%), germacrene-D (10.6%), n-octane (7.3%), α-selinene (6.5%) and β-selinene (5.5%) as main constituents. Sixty-seven components were identified in the oil of H. tomentosum with menthone (17.0%), n-octane (9.9%), β-caryophyllene (5.3%), α-pinene (5.2%), lauric acid (4.1%) and β-pinene (3.7%) as the most abundant components. Both oils were characterized by the presence of many components which could have numerous applications in food, pharmaceutical and perfume industries.     

Chemical composition and anti-fungal properties of the essential oils and crude extracts of Orthosiphon stamineus Benth

The hydrodistilled essential leaves and stems oils of Orthosiphon stamineus Benth were analysed by GC–MS/MS. Sixty nine compounds representing 97.6 and 97.4% of the total leaves and stems oils, respectively were identified, of which β-caryophyllene (24.0 and 35.1%), α-humulene (14.2 and 18.4%), β-elemene (11.1 and 8.5%), 1-octen-3-ol (8.2 and 7.0%), β-bourbonene (3.4 and 3.0%), β-pinene (2.1 and 1.7%), caryophyllene oxide (1.6 and 2.2%), camphene (1.6 and 1.3%) and limonene (1.2 and 1.1%) were the major compounds. Thus, the monoterpenes and sesquiterpenes were the predominant portions of the oils. Essential oils and methanol extract of O. stamineus and the derived fractions of hexane, chloroform, and ethyl acetate were tested for anti-fungal activity, which was determined by disc diffusion and minimum inhibitory concentration (MIC) determination methods. The oils, methanol extract and derived fractions of methanol extract displayed great potential of anti-fungal activity as a mycelial growth inhibitor against the tested phytopathogenic fungi such as Botrytis cinerea, Rhizoctonia solani, Fusarium solani, Colletotricum capsici and Phytophthora capsici, in the range of 49.3–70.3% and minimum inhibitory concentration ranging from 500 to 1000 μg/ml.     

The fermentation of trahanas: a milk-wheat flour combination

The fermented food, trahanas (a milk-wheat flour combination prepared in Greece), was studied to determine the microbiological and chemical changes that occur during fermentation. It is a lactic acid bacterial fermentation in whichStreptococcus lactis, Streptococcus diacetylactis, Leuconostoc cremoris, Lactobacillus lactis, Lactobacillus casei, Lactobacillus bulgaricus andLactobacillus acidophilus play the major acid- and aroma-producing role. The whole fermentation lasts about 50 hours. The pH of the final dried trahanas was 4.07–4.75, the acidity 0.60–1.00%, the moisture content 8.6–11.5% and the protein content 10.4–13.6%. The product offers possibilities as an increased nutritive value or high-protein food.     

The study of shea butter. VI: The extraction of shea butter

Shea butter has been extracted from the seeds of the shea tree,B. parkii, with various organic solvents. Petroleum ether (40°–60°C),n-hexane, chloroform, and benzene extracted 32%–38% of fat and 8–9 mg% of vitamin E. These solvents, particularly petroleum ether andn-hexane, can be used for the production of shea butter that is free from any oxidized fat and coloring impurities.     

Variability in essential oil and bioactive chiral monoterpenoid compositions of Indian oregano (Origanum vulgare L.) populations from northwestern Himalaya and their chemotaxonomy

The chemical composition of essential oils isolated from aerial parts of native populations of Origanum vulgare L. from northwestern Himalaya was investigated by enantioselective gas chromatography (enantio-GC), capillary-gas chromatography (capillary-GC) and GC–MS. Seventy compounds, which accounted for 84.9–97% of the oil composition, have been identified. The oils were allotted to two main groups; the first group had a high percentage of components belonging to the thymol/carvacrol biosynthetic pathway and the second was characterized by a different sesquiterpene composition and a high linalool and bornyl acetate percentage. Essential oils of O. vulgare populations from Rilkot and Dhanachuli areas in northwestern Himalaya were characterized by high monoterpenoids proportions as compared to Nainital and Bhowali populations, which were dominated by sesquiterpenoids. High proportions of thymol (29.2–82%) were found only in two populations. Composition of the samples collected from Dhanachuli area differed from Nainital and Bhowali collections by a unique presence of carvacrol. Notably, there were significant differences in the concentration of rest of the terpenes in all the collections. On enantio-GC, enantiomeric excess for (3R) (−)- and (3S) (+)-linalool, (S) (+)- and (R) (−)-terpinen-4-ol, (S) (−)- and (R) (+)-α-terpineol, (1S) (−)-bornyl acetate and (1S) (−)-borneol were established in populations from Bhowali and Dhanachuli area. Moreover, linalool type oregano may be useful in promoting wider commercial utilization of the biodiversity of this species.     

Antioxidant Effect of Dietary Supplement Withania somnifera L. Reduce Blood Glucose Levels in Alloxan-Induced Diabetic Rats

The phenolic compounds and flavonoids were determined from the extracts of Withania somnifera root (WSREt) and leaf (WSLEt). The WSREt has 28.26 mg/g total phenolic compounds and 17.32 mg/g flavonoids, whereas WSLEt has 5.4 mg/g total phenolic compounds and 5.1 mg/g flavonoids. The WSREt, WSLEt and glibenclamide were orally administered daily to diabetic rats for 8 weeks. After the treatment, the levels of urine sugar, blood glucose, liver glycogen, and antioxidants like vitamin C and E in plasma and superoxide dismutase (SOD), catalase (CAT), thiobarbituric acid reactive substances (TBARS), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and reduced glutathione (GSH) in liver, kidney and heart were determined. Diabetic rats showed a significant (p < 0.05) elevation in glucose and TBARS and a significant (p < 0.05) reduction in glycogen, vitamin C and E, SOD, CAT, GPx, GST, and GSH levels when compared to normal control rats. Administration of WSREt, WSLEt and glibenclamide to diabetic rats restored the levels to normal. In the light of aforesaid facts, it is suggested that the presence of phenolic compounds including flavonoids in W. somnifera root and leaf extracts and their antioxidant activity may play a vital role in reduction of blood glucose level in alloxan-induced diabetic rats.     

Yield, grain quality and water use efficiency of rice under non-flooded mulching cultivation

Plastic film or straw mulching cultivation under non-flooded condition has been considered as a new water-saving technique in rice production. This study aimed to investigate the yield performance in terms of quality and quantity and water use efficiency (WUE) under such practices. A field experiment across 3 years was conducted with two high-yielding rice cultivars, Zhendao 88 (a japonica cultivar) and Shanyou 63 (an indica hybrid cultivar) and four cultivation treatments imposed from transplanting to maturity: traditional flooding as control (TF), non-flooded plastic film mulching (PM), non-flooded wheat straw mulching (SM), and non-flooded no mulching (NM). Compared with those under the TF, root oxidation activity, photosynthetic rate, and activities of key enzymes in sucrose-to-starch conversion in grains during the grain filling period were significantly increased under the SM, whereas they were significantly reduced under the PM and NM treatments. Grain yield showed some reduction under all the non-flooded cultivations but differed largely among the treatments. The reduction in yield was 7.3–17.5% under the PM, 2.8–6.3% under the SM, and 39–49% under the NM. The difference in grain yield was not significant between TF and SM treatments. WUE for irrigation was increased by 314–367% under the PM, 307–321% under the SM, and 98–138% under the NM. Under the same treatment especially under non-flooded conditions, the indica hybrid cultivar showed a higher grain yield and higher WUE than the japonica cultivar. The SM significantly improved milling, appearance, and cooking qualities, whereas the PM or the NM decreased these qualities. We conclude that both PM and SM could significantly increase WUE, while the SM could also maintain a high grain yield and improve quality of rice. The SM would be a better practice than the PM in areas where water is scarce while temperature is favorable to rice growth, such as in Southeast China.     

Direct lipase-catalyzed lipophilization of chlorogenic acid from coffee pulp in supercritical carbon dioxide

The direct esterification of chlorogenic acid (5-CGA) by immobilized Candida antarctica lipase B (Novozym 435) in supercritical CO₂/t-butanol has been studied. The Taguchi approach was applied to evaluate the effects of temperature (35–55 °C), pressure (150–250 bar), t-butanol (2–10%, v/v), and the enzyme amount (10–30 mg/ml), on the ester concentration and overall conversion. Optimum reaction conditions were established at: 150 bar, 55 °C, 10% t-butanol (v/v), 20 mg/ml of lipase. Addition of 20 mg/ml of molecular sieves (3 Å) was also necessary to minimize the inhibiting effect of the increasing water concentration. Maximum conversions reached 77, 82 and 85% in 25 h using geraniol, pentanol and heptanol as aliphatic chain donors, respectively. HPLC analysis demonstrated the selective esterification of 5-CGA from a coffee pulp aqueous-methanolic extract, which reached a conversion to heptyl ester of 65% at the optimal condition. The supercritical CO₂ selectivity towards the esterified product was the working principle of this study, by which minimized interphase transport limitations and enhanced mass-transfer phenomena substantially improved the reaction kinetics.     

The cocoyam,Xanthosoma sagittifollium,as a potential raw material source for beer brewing

A widely cultivated cocoyam variety,Xanthosoma sagittifollium, was assessed for its suitability for lager beer production, using maltedSorghum vulgare (white variety) for saccharification of the substrate. The three-mash decoction method was used. Results showed that the cocoyam was superior to barley and sorghum as a substrate because of its potentially higher carbohydrate content (71–78%) compared to barley (65%) and sorghum (70–73%). The percentage wort extract was 13.3–14.5° plato compared to 7–12 for barley and 13° for the tuber crop, cassava. The kiln driedXanthosoma sagittifollium gave a dark bager beer with good aroma (68%) and very good flavour (73%) when compared with a commercial lager beer (Monarch) as standard (100%). The alcohol content (4.16% w/v) and the specific gravity (1013) were within the range (4.0–6.4%) and (1011–1019), respectively, specified by the Standard Organization of Nigeria. Although the bitterness value 40 EBU, was higher than the typical range 16–30 EBU, the taste panel indicated this was acceptable. The pH was 4.68 compared with the standard 4.36, while the acidity (as % lactic acid) was 0.36 compared to 0.24 for some commercial products.     

Occurrence and seasonal distribution of leafhopper vectors of the X-disease causal agent in methoxychlor-sprayed and unsprayed peach orchards

Leafhopper (LH) (Homoptera: Cicadellidae) populations were monitored in a peach orchard in which eight blocks of 10 trees each were unsprayed or sprayed seven times in 1979 and three times in 1980 with methoxychlor. Of 7552 LH trapped during both years, 1335 adults of six species known to transmit the mycoplasma-like causal agent of X-disease (X-vectors) were recorded: Paraphlepsius irroratus (Say), 703; Scaphytopius acutus (Say), 439; Colladonus clitellarius (Say), 88; Norvellina seminuda (Say), 65; Gyponana lamina (Delong), 39; and Fieberiella florii (Stål), three. In sprayed blocks, total LH, total X-vectors and total P. irroratus and S. acutus adults (the most abundant X-vectors), were reduced by 49%, 46%, 43% and 50% respectively. In 1979, during one of two periods of peak adult LH activity, total LH, total X-vectors, P. irroratus and S. acutus were reduced by 56–73%, 22–48%, 26% and 62–85% respectively. In 1980, during the first period of adult leafhopper activity, total LH, total X-vectors, P. irroratus and S. acutus populations in sprayed trees were reduced by 72%, 56%, 50% and 60% respectively. The value of reducing X-vector populations, especially those of major X-vectors such as S. acutus and P. irroratus, for disease control is unknown, but should be considered for integrated management of X-disease.     

Promotive effects of long-term fertilization on growth of tissue culture-derived Hypericum polyanthemum plants during acclimatization

The effect of long-term fertilization supply on biomass yield and secondary metabolites accumulation was assessed in vegetative and reproductive parts of acclimatized field grown plants of Hypericum polyanthemum, an endemic species of southern Brazil. Fertilization caused positive responses in plant growth with biomass increment of vegetative and reproductive parts, proportional to the MS solution concentration (25%, 50% and 100%), whereas the chemical analyses indicated that the supply of fertilization solution did not alter the concentration of 6-isobutyryl-5,7-dimethoxy-2,2-dimethyl-benzopyran (HP1), 7-hydroxy-6-isobutyryl-5-methoxy-2,2-dimethyl-benzopyran (HP2), uliginosin B as well as total phenolic compounds (TPCs), and increased the accumulation of 5-hydroxy-6-isobutyryl-7-methoxy-2,2-dimethyl-benzopyran (HP3) in all fertilization regimes.     

Differences in conformational stability of barley alpha-amylase isozymes 1 and 2. Role of charged groups and isozyme 2 specific salt-bridges

Barley alpha-amylase isozymes 1 (AMY1) and 2 (AMY2) have 80% sequence identity but possess different physico-chemical properties. By incubation in the range 37–85 °C T₅₀ is 75.2 °C of AMY1 and 79.2 °C of AMY2. While AMY2 is also most stable in urea at pH 6.7, [urea]₅₀ being 8.2 M compared to 7.9 M for AMY1, AMY1 has highest stability in urea below pH 6 or in the presence of NaCl. Moreover AMY1 is most stable in guanidinium chloride. Charge screening thus destabilises AMY2 but stabilises AMY1. Isozyme sequence comparison suggests that AMY1 lacks four of the 20 salt-bridges identified in the crystal structure of AMY2. The four residues that differ comprise Lys67_AMY2 and Asp267_AMY2, forming salt-bridges on the surface of the catalytic (β/α)₈-barrel (domain A), and Glu96_AMY2 and His344_AMY2 that participate in charged networks between domain A and the small domain B and the C-terminal domain, respectively. Four corresponding AMY2 mimics A68K; D97E; Q269D; N346H were made in AMY1 by site-directed mutagenesis. While D97E and Q269D have slightly improved stability compared to AMY1 wild-type, N346H and, under certain conditions, A68K are destabilised. The four mutants show 22–176% activity (k_cat/K_m) toward 2-chloro-4-nitrophenol β- -maltoheptaoside and amylose DP17 and 43–117% activity for insoluble starch.     

Efficacy of Rheum officinale liquid formulation on cucumber powdery mildew

Rheum officinale liquid formulation, the ethanol extract from roots of R. officinale Baill., formulated as physcion 5 g l⁻¹ aqueous solution (AS), has been commercialized in China for controlling cucumber powdery mildew (Podosphaera xanthii (Castagne) U. Braun & Shishkoff). The efficacy of the product was evaluated in pot tests under controlled conditions and in open and protected fields in China over 2 years. In most trials, the efficacy reached above 80% at the rates of 10–50 mg a.i l⁻¹ water after three applications and at the rates of 20–50 mg a.i l⁻¹ water after two applications. The cucumber fruit yield in the product treatment was as many or more as triadimefon treatment, and over 53.1% compared to the untreated control. These results showed that Chinese rhubarb extract could be an effective alternative plant protecting agent in the integrated and biological management of cucumber powdery mildew.